| Project/Area Number |
16580236
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Basic veterinary science/Basic zootechnical science
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| Research Institution | Health Sciences University of Hokkaido (2005)
Hokkaido University (2004) |
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Principal Investigator |
OKAZAKI Katsunori Health Sciences University of Hokkaido, Dept of Pharmaceutical Sciences, Professor, 薬学部, 教授 (90160663)
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| Co-Investigator(Kenkyū-buntansha) |
KIDA Hiroshi Hokkaido University, Graduate School of Veterinary Medicine, Professor, 大学院・獣医学研究科, 教授 (10109506)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 2005: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 2004: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | Virus envelope / Membrane fusion / Cell entry / Herpesviru / gB glycoprotein / Proteolytic enzyme / Endothelial cells / Endosome / ヘプタドリピート / gB / 糖蛋白 / フリン |
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| Research Abstract |
Glycoprotein B (gB) is the most conserved glycoprotein of herpesviruses and plays important roles in virus infectivity. The glycoprotein of most herpesviruses, including psuedorabies virus (PrV), is cleaved by a cellular protease into two disulfide-linked subunits. In the present study, we demonstrated that the glycoprotein generated in human colon carcinoma Lo Vo cells, which lack the ubiquitous protease furin, remained the uncleaved form and that the virus replicated in Lo Vo cells without cell fusion. The uncleaved gB was converted into the subunits after furin digestion. The virus also replicated in Madin-Darby bovine kidney cells in the presence of a peptide furin inhibitor without cell fusion although distinguished syncytia were formed in the absence of the inhibitor. Lo Vo cells constitutively expressing furin showed cell fusion when they were infected with the virus. The penetration kinetics assays revealed that the virus carrying uncleaved gB penetrated the cells at the same rate as that carrying cleaved gB. These results indicate that PrV gB was cleaved by furin and that the cleavage was dispensable for virus replication in vitro. gB cleavage was involved in syncytium formation but not in penetration kinetics, suggesting that different mechanisms operated between cell-cell fusion and virus-cell fusion by PrV.
Electron microscopic examination at the stage of entry of equine herpesvirus 1 (EHV-1) showed that viral particles were present within uncoated vesicles in the cytoplasm of equine brain microvascular endothelial cells (EBMECs). Although herpesviruses are thought to enter the cells on the plasma membrane, these results might show that EHV-1 enter EBMECs through endosome.
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